DOI: 10.1046/j.1365-313x.1992.t01-41-00999.x. Vicilin with carboxy‐terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants

Vicilin with carboxy‐terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants

10.1046/j.1365-313x.1992.t01-41-00999.x

Crossref journal-article

Wiley

The Plant Journal (311)

Abstract

SummaryGene constructs were designed to test the effect of the endoplasmic reticulum (ER)‐targeting signal, KDEL, on the level of accumulation of a foreign protein in transgenic plants. The gene for the pea seed protein vicilin was modified by the addition of a sequence coding for this tetrapeptide at its carboxyl terminus. The altered gene was placed under the control of a CaMV 35S promoter and its expression in the leaves of both tobacco and lucerne (alfalfa) was compared with that of an equivalent vicilin construct lacking the KDEL‐coding sequence. The presence of the ER‐targeting signal led to a greatly enhanced accumulation of the heterologous protein. In lucerne and tobacco leaves, the level of vicilin‐KDEL protein was 20 and 100 times greater than that of the unmodified vicilin, respectively. These differences in expression level could not be explained by corresponding differences in the steady‐state levels or the translatability of the mRNAs. However, when the stability of vicilin and vicilin‐KDEL proteins was compared in their respective transgenic hosts, unmodified vicilin was found to be degraded with a half‐life of 4.5 h while vicilin‐KDEL was much more stable with a half‐life of more than 48 h.Immunogold labelling of leaf tissues from transgenic lucerne and tobacco showed the presence of vicilin associated with large aggregates within the ER lumen of vicilin‐KDEL plants. No such aggregates were detected in transgenic plants expressing wild‐type vicilin.It is concluded that the carboxy‐terminal KDEL caused the retention of the modified vicilin in the ER, and that this retention led to the increased stability and higher level of accumulation of vicilin‐KDEL in leaves of transgenic plants.

Bibliography

Wandelt, C. I., Khan, M. R. I., Craig, S., Schroeder, H. E., Spencer, D., & Higgins, T. J. V. (1992). Vicilin with carboxyâterminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants. The Plant Journal, 2(2), 181â192. Portico.

Authors 6

Christine I. Wandelt (first)
M. Rafiqul I. Khan (additional)
Stuart Craig (additional)
Harmut E. Schroeder (additional)
Donald Spencer (additional)
Thomas J.V. Higgins (additional)

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10.1083/jcb.109.6.2633

Dates

Type	When
Created	22 years, 5 months ago (March 12, 2003, 4:39 p.m.)
Deposited	1 year, 10 months ago (Oct. 23, 2023, 11:10 a.m.)
Indexed	1 month, 2 weeks ago (July 20, 2025, 12:03 a.m.)
Issued	33 years, 6 months ago (March 1, 1992)
Published	33 years, 6 months ago (March 1, 1992)
Published Online	20 years, 6 months ago (Feb. 23, 2005)
Published Print	33 years, 6 months ago (March 1, 1992)

Funders 0

None

BibTeX

@article{Wandelt_1992, title={Vicilin with carboxy‐terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants}, volume={2}, ISSN={1365-313X}, url={http://dx.doi.org/10.1046/j.1365-313x.1992.t01-41-00999.x}, DOI={10.1046/j.1365-313x.1992.t01-41-00999.x}, number={2}, journal={The Plant Journal}, publisher={Wiley}, author={Wandelt, Christine I. and Khan, M. Rafiqul I. and Craig, Stuart and Schroeder, Harmut E. and Spencer, Donald and Higgins, Thomas J.V.}, year={1992}, month=mar, pages={181–192} }

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